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lüll Vigilance and validation: Keys to success in RNAi screening Sigoillot FD; King RWACS Chem Biol 2011[Jan]; 6 (1): 47-60In the 12 years since the process of RNA interference (RNAi) was first discovered, great progress has been made in understanding its mechanism and exploiting its ability to silence gene expression to study gene function at a genome-wide level. Its extensive use as a screening method has yielded many published lists of genes that play novel roles in higher eukaryotes. However, the usefulness of this information is potentially limited by the occurrence of unintended off-target effects. Here we review the potential causes of off-target effects and the impact of this phenomenon in interpreting the results of high-throughput RNAi screens. In addition to targeting the intended gene product, artificial short interfering RNAs (siRNAs) can produce off-target effects by down-regulating the expression of multiple mRNAs through microRNA-like targeting of the 3' untranslated region. We examine why this phenomenon can produce high hit rates in siRNA screens and why independent validation of screening results is critical for the approach to yield new biological insights.|*RNA Interference[MESH]|Down-Regulation[MESH]|Gene Expression[MESH]|Genetic Techniques/*standards[MESH]|Genomics[MESH]|MicroRNAs/*genetics[MESH]|RNA, Double-Stranded/*chemistry[MESH]|RNA, Messenger/*genetics[MESH]|RNA, Small Interfering/*genetics/metabolism[MESH]|Transcription, Genetic[MESH]|Untranslated Regions[MESH]|Validation Studies as Topic[MESH] |